Influence of urokinase gene knockout on tissue levels of biogenic amines in mice with melanoma.

e22109 Background: The plasminogen activation system and biogenic amines are involved in melanoma pathogenesis, but currently, the mutual influence of the components of these systems on each other is unknown. The purpose of the study was to reveal the dynamics of biogenic amines in the brain and intact and tumorous skin (B16/F10 melanoma) in C57BL/6-PlautmI.IBug-ThisPlau6FDhu/GFDhu mice with uPA gene knockout. Methods: The study included male and female C57BL/6-PlautmI.IBug-This Plau6FDhu/GFDhu mice, n = 38; the comparison group included C57BL/6 mice without uPA gene knockout, n = 61. Melanoma was transplanted under the skin of the back. Levels of biogenic amines were measured in tissues taken in week 3 of carcinogenesis by ELISA using standard test systems (Cusabio, China). Results: The total content of biogenic amines was elevated in tissues of intact C57BL/6-PlautmI.IBug-ThisPlau6FDhu/GFDhu mice: in the skin – due to the noradrenaline (NA) rise by 4.8 times in males and by 4.9 times in females, histamine (H) – by 3.6 times in males and 1.6 times (p < 0.05) in females, serotonin (5HT) by 3.4 times in males and 8.3 times in females; in the brain – due to the NA rise by 3.5 times in males and 3.2 times in females, dophamine by 2.1 times in males and 2.9 times in females, while H levels declined. Distinctive features of the melanoma development in C57BL/6-PlautmI.IBug-ThisPlau6FDhu/GFDhu mice included: lower levels of adrenaline with high NA concentrations and an enhanced synthesis and inactivation of 5HTin the brain. The content of catecholamines in the tumor nodes was the same as in C57BL/6 mice, with a high concentration of H recorded in the tumor and skin, together with a high 5HT level in the skin. Conclusions: The uPA gene knockout limits the development of stress at the central regulatory and peripheral effector levels, and modulates the immune antitumor response by increasing serotonergic mediation in the brain and increasing serotonin and histamine levels in the skin of mice with B16/F10 melanoma.

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Specificity of aminergic status in skin and tumor of C57BL/6-PlautmI.IBug-This Plau6FDhu/GFDhu mice with B16/F10 melanoma

Problems in oncology ◽

10.37469/0507-3758-2020-66-6-707-711 ◽

2020 ◽

Vol 66 (6) ◽

pp. 707-711

Author(s):

Oleg Kit ◽

Elena Frantsiyants ◽

Irina Kaplieva ◽

Ekaterina Surikova ◽

Irina Neskubina ◽

...

Keyword(s):

Biogenic Amines ◽

Knockout Mice ◽

Gene Knockout ◽

Local Stress ◽

Plasminogen Activation ◽

Intact Skin ◽

Gene Knockout Mice ◽

Hydroxyindoleacetic Acid ◽

Solitary Metastases ◽

Deficient Mice

Background. Systems of plasminogen activation and biogenic amines are involved in carcinogenesis, but their relationship has not been established. Aim – study of the quantitative specificity of biogenic amines in the skin and melanoma in urokinase-gene knockout mice. Materials and methods. Levels of catecholamines, histamine, serotonin and 5-hydroxyindoleacetic acid (HIAA) were determined by ELISA in the skin and В16/F10 melanoma of urokinase (uPA) -gene knockout mice of both genders (n=24); С57ВL/6 mice (n=64) were controls (C). Results. Differential characteristics of melanoma development in uPA-deficient mice included: an earlier onset of the primary tumor and its slow growth, more pronounced in females, in combination with hemorrhages in the lungs of males and solitary metastases in the lungs of females. This was facilitated by higher levels of norepinephrine in the skin of males/females – by 4.8/4.9 times, histamine – by 3.6/1.7 (p<0.05) times and serotonin – by 3.4/8.3 times. Dopamine accumulated in melanoma in all uPA-gene knockout mice: in females –1.6 times (p<0.05), in males – 2.1 times higher than in intact skin, with a 2.5 times reduction of norepinephrine in females. Levels of histamine decreased, but exceeded controls: in females – by 1.8 times (p<0.05), in males –by 3.5 times. Levels of serotonin in uPA-deficient females were as high, while in males they were 3.4 times lower than in intact skin. Conclusions. The specificity of the aminergic system in the skin of uPA-gene knockout mice demonstrated the inhibition of local stress and contributed to the reduction of malignant potential of melanoma by increasing immune properties of the skin.

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Aminergic status in the brain of urokinase gene-deficient mice with malignant tumors growing in presence of chronic neurogenic pain.

Journal of Clinical Oncology ◽

10.1200/jco.2021.39.15_suppl.e21582 ◽

2021 ◽

Vol 39 (15_suppl) ◽

pp. e21582-e21582

Author(s):

Yulia A. Pogorelova ◽

Irina V. Kaplieva ◽

Elena M. Frantsiyants ◽

Ekaterina I. Surikova ◽

Valeria A. Bandovkina ◽

...

Keyword(s):

Sciatic Nerve ◽

Tumor Growth ◽

Biogenic Amines ◽

Malignant Tumors ◽

Gene Knockout ◽

Significant Inhibition ◽

Mouse Strains ◽

Neurogenic Pain ◽

The Brain ◽

Melanoma Growth

e21582 Background: The fibrinolytic system of the brain is important for its normal functioning and participation in processes that are significant in various stressful influences, including tumor growth and chronic neurogenic pain (CNP). These pathological conditions change the activity of the brain neurotransmitter system. On the other hand, urokinase deficiency is associated with significant inhibition of tumor growth, while CNP – with its stimulation. The purpose of the study was to analyze the effect of CNP on the levels of biogenic amines in the brain of mice with urokinase deficiency (uPA-/-) with transplanted B16/F10 melanoma. Methods: The study included male and female mice: С57ВL/6 (uPA+/+, n = 48) and C57BL/6-Plautm1.1Bug-ThisPlauGFDhu/GFDhu (urokinase gene-knockout - uPA-/-, n = 48). Mouse strains were divided into subgroups (each n = 6): intact; with CNP (bilateral sciatic nerve ligation); 21 days after subcutaneous transplantation of B16/F10 melanoma; 21 days of B16/F10 melanoma growth in presence of CNP (B16/F10+CNP), with tumor transplantation 2 weeks after the sciatic nerve ligation. Levels of adrenaline (A), noradrenaline (NA), dopamine (DA), histamine, serotonin (5HT), and 5-hydroxyindoleacetic acid (5OHIA) were determined in the brain by ELISA (Cusabio, China). Statistical processing - Statistica 10.0. Results: Levels of NA, DA and 5HT in the brain of intact uPA-/- mice were 3.5, 2.1 and 1.9 times higher (p < 0.05), respectively, than in intact uPA+/+ animals, while histamine and 5OHIA were on average 2.0 times lower. The dynamics of cerebral levels of biogenic amines in uPA-/- mice with pathological factors, alone or combined, had practically no gender specificity, with rare exceptions. So, 5HT levels increased up to 4.5 times in uPA-/- mice of both sexes in response to CNP or B16/F10 growth. Melanoma growth in presence of CNP, on the contrary, decreased 5HT by 3-10 times and DA by 1.6 times (p < 0.05) both in males and females, and decreased NA by 1.6 times (p < 0.05) in females. Conclusions: CNP together with melanoma inhibits the initial activation of the HA-, DA- and 5HT-ergic systems in the brain of uPA-/- mice, which may be an important pathogenetic mechanism of the cancellation of genetically determined inhibition of subcutaneous B16/F10 melanoma growth in urokinase deficiency.

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Factors Involved In the Plasminogen Activation System in Human Breast Tumours

Thrombosis and Haemostasis ◽

10.1055/s-0038-1642505 ◽

1994 ◽

Vol 71 (05) ◽

pp. 684-691 ◽

Cited By ~ 9

Author(s):

László Damjanovich ◽

Csaba Turzó ◽

Róza Ádány

Keyword(s):

Epithelial Cells ◽

Connective Tissue ◽

Plasminogen Activators ◽

Breast Tumour ◽

Plasminogen Activation ◽

Malignant Tumours ◽

Plasminogen Activation System ◽

Activation System ◽

Malignant Breast ◽

Pai 1

SummaryThe plasminogen activation system is a delicately balanced assembly of enzymes which seems to have primary influence on tumour progression. The conversion of plasminogen into serine protease plasmin with fibrinolytic activity depends on the actual balance between plasminogen activators (urokinase type; u-PA and tissue type; t-PA) and their inhibitors (type 1 and 2 plasminogen activator inhibitors; PAI-1 and PAI-2). The purpose of this study was to determine the exact histological localization of all the major factors involved in plasminogen activation, and activation inhibition (plasmin system) in benign and malignant breast tumour samples. Our results show that factors of the plasmin system are present both in benign and malignant tumours. Cancer cells strongly labelled for both u-PA and t-PA, but epithelial cells of fibroadenoma samples were also stained for plasminogen activators at least as intensively as tumour cells in cancerous tissues. In fibroadenomas, all the epithelial cells were labelled for PAM. Staining became sporadic in malignant tumours, cells located at the periphery of tumour cell clusters regularly did not show reaction for PAI-1. In the benign tumour samples the perialveolar connective tissue stroma contained a lot of PAI-1 positive cells, showing characteristics of fibroblasts; but their number was strongly decreased in the stroma of malignant tumours. These findings indicate that the higher level of u-PA antigen, detected in malignant breast tumour samples by biochemical techniques, does not necessarily indicate increased u-PA production by tumour cells but it might be owing to the increased number of cells producing u-PA as well. In malignant tumours PAI-1 seems to be decreased in the frontage of malignant cell invasion; i.e. malignant cells at the host/tumour interface do not express PAI-1 in morphologically detectable quantity and in the peritumoural connective tissue the number of fibroblasts containing PAI-1 is also decreased.

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Interaction of Heparin with Fibrinolysis

Thrombosis and Haemostasis ◽

10.1055/s-0038-1654997 ◽

1963 ◽

Vol 09 (02) ◽

pp. 446-458 ◽

Cited By ~ 5

Author(s):

Rudolf Holemans ◽

Dionysios Adamis ◽

James F Horace

Keyword(s):

Human Plasma ◽

Fibrinolytic Activity ◽

Plasminogen Activation ◽

Fibrinolytic Agents ◽

High Concentration ◽

Enhancing Effect

SummaryHeparin in high concentration inhibits the fibrinolysis of human plasma clots or bovine fibrin by fibrinolytic agents which produce plasminogen activation. Heparin has no effect on the fibrinolytic activity of plasmin or Aspergillus protease.In order to produce inhibition of plasminogen activation heparin requires the presence of a co-factor which is present in citrated human plasma but absent from its euglobulin fraction.In none of the concentrations tested has heparin an enhancing effect on fibrinolysis.

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Tissue plasminogen activator is a ligand of cation-independent mannose 6-phosphate receptor and consists of glycoforms that contain mannose 6-phosphate

Scientific Reports ◽

10.1038/s41598-021-87579-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

James J. Miller ◽

Richard N. Bohnsack ◽

Linda J. Olson ◽

Mayumi Ishihara ◽

Kazuhiro Aoki ◽

...

Keyword(s):

Plasminogen Activator ◽

Tissue Plasminogen Activator ◽

Dependent Manner ◽

Plasminogen Activation ◽

Urokinase Plasminogen Activator Receptor ◽

Plasminogen Activation System ◽

Extracellular Region ◽

Glycosylation Sites ◽

Tissue Plasminogen ◽

Mannose 6 Phosphate

AbstractPlasmin is the key enzyme in fibrinolysis. Upon interaction with plasminogen activators, the zymogen plasminogen is converted to active plasmin. Some studies indicate plasminogen activation is regulated by cation-independent mannose 6-phosphate receptor (CI-MPR), a protein that facilitates lysosomal enzyme trafficking and insulin-like growth factor 2 downregulation. Plasminogen regulation may be accomplished by CI-MPR binding to plasminogen or urokinase plasminogen activator receptor. We asked whether other members of the plasminogen activation system, such as tissue plasminogen activator (tPA), also interact with CI-MPR. Because tPA is a glycoprotein with three N-linked glycosylation sites, we hypothesized that tPA contains mannose 6-phosphate (M6P) and binds CI-MPR in a M6P-dependent manner. Using surface plasmon resonance, we found that two sources of tPA bound the extracellular region of human and bovine CI-MPR with low-mid nanomolar affinities. Binding was partially inhibited with phosphatase treatment or M6P. Subsequent studies revealed that the five N-terminal domains of CI-MPR were sufficient for tPA binding, and this interaction was also partially mediated by M6P. The three glycosylation sites of tPA were analyzed by mass spectrometry, and glycoforms containing M6P and M6P-N-acetylglucosamine were identified at position N448 of tPA. In summary, we found that tPA contains M6P and is a CI-MPR ligand.

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THE HYDROLYSIS OF MONOPHOSPHOINOSITIDE BY EXTRACTS OF BRAIN

Canadian Journal of Biochemistry ◽

10.1139/o67-095 ◽

1967 ◽

Vol 45 (6) ◽

pp. 853-861 ◽

Cited By ~ 60

Author(s):

W. Thompson

Keyword(s):

Fatty Acids ◽

Enzyme Activity ◽

Calcium Ions ◽

Brain Extract ◽

Monovalent Cations ◽

High Concentration ◽

Diffusible Factor ◽

Hydrolysis Of ◽

The Brain ◽

Long Chain

The hydrolysis of monophosphoinositide by soluble extracts from rat brain is described. Diglyceride and inositol monophosphate are liberated along with a small amount of free fatty acids. Hydrolysis of the lipid is optimal at pH 5.4 in acetate buffer. The reaction is stimulated by calcium ions or by high concentration of monovalent cations and, to a less extent, by long-chain cationic amphipathic compounds. Enzyme activity is lost on dialysis of the brain extract and can be restored by diffusible factor(s). Some differences in the conditions for hydrolysis of mono- and tri-phosphoinositides are noted.

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